Immune challenged SC-islets first response profiled by single cell transcription analysis after co-culture with human allogeneic PBMCs [scRNA-seq_invitro]

We performed an in vitro co-culture of allogeneic PBMCs and SC-islet clusters. SC-islet clusters were enriched for ß cells (using CD49A magnetic sorting; SC-a still remain at lower numbers) (Veres et al., 2019), dissociated and reaggregated to obtain a more uniform cell count between wells. SC-islets were co-cultured with human allogeneic PBMCs for 24 or 48 hours. As controls [time(t)=0], SC-islets remained in culture without PBMC addition. These samples, in addition to PBMCs alone (t=0), were used for scRNA-seq (Figure 2A). Overall design: Comparing expression datasets from PBMC co-cultured with SC-islets, to SC-islets alone

我们开展了同种异体外周血单个核细胞（peripheral blood mononuclear cells, PBMCs）与干细胞来源胰岛类器官簇（stem cell-derived islet clusters, SC-islet clusters）的体外共培养实验。该SC-islet簇经CD49A磁珠分选富集了β细胞（SC-α细胞仍维持较低丰度）（Veres等，2019），随后将其解离并重新聚集，以确保各培养孔间的细胞计数更为均一。将SC-islet与人类同种异体PBMCs共培养24小时或48小时。以未添加PBMCs的SC-islet培养体系作为对照组（时间点t=0）；此外单独培养的PBMCs（t=0）样本均用于单细胞RNA测序（single-cell RNA sequencing, scRNA-seq），实验设计详见图2A。整体实验设计为：比较与SC-islet共培养的PBMCs的基因表达数据集与单独培养的SC-islet的基因表达数据集之间的差异。